| 质粒名称: | pHSN6I01 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 50587 |
| 存储实验室: | Qi-Jun Chen |
|
载体骨架基本信息 |
|
| 载体名称: | pGreen-like binary vector |
| 载体出品公司: | Mullineaux Lab, see http://www.pgreen.ac.uk/ |
| 载体抗性: | 卡那霉素和大观霉素 |
| 载体类型: | CRISPR ;植物表达 |
| 筛选标记: | 潮霉素 |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | dCas9-KRAB,Cas9 |
| 物种来源: | Synthetic |
| 基因突变: | dCas9 that is defective in DNA cleavage; the maize codon–optimized dCas9-KRAB (Krüppel-associated box) vector was constructed for targeted knockdown of genes of interest. |
| 启动子 | 2×35Sp |
| 标签/融合蛋白: |
3xFLAG（插入件上的N端子）
NLS（插入件上的N端子）
NLS（插入件上的C端子）
HA（插入件上的C端子）
|
| 5' 测序引物 | hSpCas9-R1, CGCTCGTGCTTCTTATCCTC |
|
文献引用方法 |
|
| 材料和方法部分: | pHSN6I01 was a gift from Qi-Jun Chen (Addgene plasmid # 50587 ; http://n2t.net/addgene:50587 ; RRID:Addgene_50587) |
| 参考文献部分: | A CRISPR/Cas9 toolkit for multiplex genome editing in plants. Xing HL, Dong L, Wang ZP, Zhang HY, Han CY, Liu B, Wang XC, Chen QJ. BMC Plant Biol. 2014 Nov 29;14(1):327. 10.1186/s12870-014-0327-y PubMed 25432517 |
买家导航
载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS pHSN6I01 13088 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION CRISPR/Cas based plant genome editing and gene regulation; expresses
dCas9-KRAB, gRNA scaffold for insertion of target sequence (AtU6-26
promoter), Hyg resistance.
ACCESSION .
VERSION .
KEYWORDS pHSN6I01
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 13088)
AUTHORS Xing HL, Dong L, Wang ZP, Zhang HY, Han CY, Liu B, Wang XC, Chen QJ
TITLE A CRISPR/Cas9 toolkit for multiplex genome editing in plants.
JOURNAL BMC Plant Biol. 2014 Nov 29;14(1):327.
PUBMED 25432517
REFERENCE 2 (bases 1 to 13088)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..13088
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_feature 1..25
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
CDS 810..1601
/codon_start=1
/gene="aadA"
/product="aminoglycoside adenylyltransferase (Murphy,
1985)"
/label=SmR
/note="confers resistance to spectinomycin and
streptomycin"
/translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
misc_RNA 1753..1828
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
promoter 2442..2786
/label=CaMV 35S promoter
/note="strong constitutive promoter from cauliflower mosaic
virus"
primer_bind 2730..2750
/label=35S promoter
/note="CaMV 35S promoter, forward primer"
CDS 2969..3034
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
/label=3xFLAG
/translation="DYKDHDGDYKDHDIDYKDDDDK"
CDS 3041..3061
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 3086..7186
/codon_start=1
/product="catalytically dead mutant of the Cas9
endonuclease from the Streptococcus pyogenes Type II
CRISPR/Cas system"
/label=dCas9
/note="RNA-guided DNA-binding protein that lacks
endonuclease activity due to the D10A mutation in the RuvC
catalytic domain and the H840A mutation in the HNH
catalytic domain"
/translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
ATLIHQSITGLYETRIDLSQLGGD"
RBS 7099..7107
/label=Shine-Dalgarno sequence
/note="full consensus sequence for ribosome-binding sites
upstream of start codons in E. coli; complementary to a
region in the 3' end of the 16S rRNA (Chen et al., 1994)"
CDS 7187..7234
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
CDS 7253..7273
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 7289..7309
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 7346..7531
/codon_start=1
/gene="ZNF10"
/gene_synonym="KOX1"
/product="Kruppel-associated box (KRAB) transcriptional
repression domain from the human zinc finger protein ZNF10
(Margolin et al., 1994)"
/label=KRAB
/translation="RTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGY
QLTKPDVILRLEKGEEP"
CDS 7538..7558
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 7565..7591
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
terminator 7644..7896
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
promoter 8012..8689
/label=CaMV 35S promoter (enhanced)
/note="cauliflower mosaic virus 35S promoter with a
duplicated enhancer region"
CDS 8757..9782
/codon_start=1
/gene="aph(4)-Ia"
/product="aminoglycoside phosphotransferase from E. coli"
/label=HygR
/note="confers resistance to hygromycin"
/translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
K"
polyA_signal 9822..9996
/label=CaMV poly(A) signal
/note="cauliflower mosaic virus polyadenylation signal"
misc_feature 10146..10170
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
primer_bind complement(10418..10441)
/label=pENTR-F
/note="pENTR vectors, forward primer"
terminator 10449..10476
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator 10568..10654
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
rep_origin 10823..11258
/label=pSa ori
/note="origin of replication from bacterial plasmid pSa"
rep_origin complement(11373..11961)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(11453..11472)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(12051..12860)
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
FQVLEEYPDSGENIVDALAVSLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
primer_bind 12774..12793
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
primer_bind 12852..12871
/label=pENTR-R
/note="pENTR vectors, reverse primer"
ORIGIN
1 gtttacccgc caatatatcc tgtcaaacac tgatagttta aactgaaggc gggaaacgac
61 aatctgatcc aagctcaagc taagctcacg tgacggaatt aagcttcgac ttgccttccg
121 cacaatacat catttcttct tagctttttt tcttcttctt cgttcataca gttttttttt
181 gtttatcagc ttacattttc ttgaaccgta gctttcgttt tcttcttttt aactttccat
241 tcggagtttt tgtatcttgt ttcatagttt gtcccaggat tagaatgatt aggcatcgaa
301 ccttcaagaa tttgattgaa taaaacatct tcattcttaa gatatgaaga taatcttcaa
361 aaggcccctg ggaatctgaa agaagagaag caggcccatt tatatgggaa agaacaatag
421 tatttcttat ataggcccat ttaagttgaa aacaatcttc aaaagtccca catcgcttag
481 ataagaaaac gaagctgagt ttatatacag ctagagtcga agtagtgatt gggagaccaa
541 cccagtggac ataagcctgt tcggttcgta agctgtaatg caagtagcgt atgcgctcac
601 gcaactggtc cagaaccttg accgaacgca gcggtggtaa cggcgcagtg gcggttttca
661 tggcttgtta tgactgtttt tttggggtac agtctatgcc tcgggcatcc aagcagcaag
721 cgcgttacgc cgtgggtcga tgtttgatgt tatggagcag caacgatgtt acgcagcagg
781 gcagtcgccc taaaacaaag ttaaacatca tgggggaagc ggtgatcgcc gaagtatcga
841 ctcaactatc agaggtagtt ggcgtcatcg agcgccatct cgaaccgacg ttgctggccg
901 tacatttgta cggctccgca gtggatggcg gcctgaagcc acacagtgat attgatttgc
961 tggttacggt gaccgtaagg cttgatgaaa caacgcggcg agctttgatc aacgaccttt
1021 tggaaacttc ggcttcccct ggagagagcg agattctccg cgctgtagaa gtcaccattg
1081 ttgtgcacga cgacatcatt ccgtggcgtt atccagctaa gcgcgaactg caatttggag
1141 aatggcagcg caatgacatt cttgcaggta tcttcgagcc agccacgatc gacattgatc
1201 tggctatctt gctgacaaaa gcaagagaac atagcgttgc cttggtaggt ccagcggcgg
1261 aggaactctt tgatccggtt cctgaacagg atctatttga ggcgctaaat gaaaccttaa
1321 cgctatggaa ctcgccgccc gactgggctg gcgatgagcg aaatgtagtg cttacgttgt
1381 cccgcatttg gtacagcgca gtaaccggca aaatcgcgcc gaaggatgtc gctgccgact
1441 gggcaatgga gcgcctgccg gcccagtatc agcccgtcat acttgaagct agacaggctt
1501 atcttggaca agaagaagat cgcttggcct cgcgcgcaga tcagttggaa gaatttgtcc
1561 actacgtgaa aggcgagatc accaaggtag tcggcaaata atgtctagct agaaattcgt
1621 tcaagccgac gccgcttcgc ggcgcggctt aactcaagcg ttagatgcac taagcacata
1681 attgctcaca gccaaactat caggtcaagt ctgcttttat tatttttaag cgtgcataat
1741 aagccggtct cggttttaga gctagaaata gcaagttaaa ataaggctag tccgttatca
1801 acttgaaaaa gtggcaccga gtcggtgctt ttttttgcaa aattttccag atcgatttct
1861 tcttcctctg ttcttcggcg ttcaatttct ggggttttct cttcgttttc tgtaactgaa
1921 acctaaaatt tgacctaaaa aaaatctcaa ataatatgat tcagtggttt tgtacttttc
1981 agttagttga gttttgcagt tccgatgaga taaaccaata agcttgcatg cctgcaggtc
2041 aacatggtgg agcacgacac acttgtctac tccaaaaata tcaaagatac agtctcagaa
2101 gaccaaaggg caattgagac ttttcaacaa agggtaatat ccggaaacct cctcggattc
2161 cattgcccag ctatctgtca ctttattgtg aagatagtgg aaaaggaagg tggctcctac
2221 aaatgccatc attgcgataa aggaaaggcc atcgttgaag atgcctctgc cgacagtggt
2281 cccaaagatg gacccccacc cacgaggagc atcgtggaaa aagaagacgt tccaaccacg
2341 tcttcaaagc aagtggattg atgtgataac atggtggagc acgacacact tgtctactcc
2401 aaaaatatca aagatacagt ctcagaagac caaagggcaa ttgagacttt tcaacaaagg
2461 gtaatatccg gaaacctcct cggattccat tgcccagcta tctgtcactt tattgtgaag
2521 atagtggaaa aggaaggtgg ctcctacaaa tgccatcatt gcgataaagg aaaggccatc
2581 gttgaagatg cctctgccga cagtggtccc aaagatggac ccccacccac gaggagcatc
2641 gtggaaaaag aagacgttcc aaccacgtct tcaaagcaag tggattgatg tgatatctcc
2701 actgacgtaa gggatgacgc acaatcccac tatccttcgc aagacccttc ctctatataa
2761 ggaagttcat ttcatttgga gaggacctcg acctcaacac aacatataca aaacaaacga
2821 atctcaagca atcaagcatt ctacttctat tgcagcaatt taaatcattt cttttaaagc
2881 aaaagcaatt ttctgaaaat tttcaccatt tacgaacgat actcgagggg gatccccaat
2941 acttgtatgg ccgcggccgc tctagatgga ttacaaggac cacgacgggg attacaagga
3001 ccacgacatt gattacaagg atgatgatga caagatggct ccgaagaaga agaggaaggt
3061 tggcatccac ggggtgccag ctgctgacaa gaagtactcg atcggcctcg ccattgggac
3121 taactctgtt ggctgggccg tgatcaccga cgagtacaag gtgccctcaa agaagttcaa
3181 ggtcctgggc aacaccgatc ggcattccat caagaagaat ctcattggcg ctctcctgtt
3241 cgacagcggc gagacggctg aggctacgcg gctcaagcgc accgcccgca ggcggtacac
3301 gcgcaggaag aatcgcatct gctacctgca ggagattttc tccaacgaga tggcgaaggt
3361 tgacgattct ttcttccaca ggctggagga gtcattcctc gtggaggagg ataagaagca
3421 cgagcggcat ccaatcttcg gcaacattgt cgacgaggtt gcctaccacg agaagtaccc
3481 tacgatctac catctgcgga agaagctcgt ggactccaca gataaggcgg acctccgcct
3541 gatctacctc gctctggccc acatgattaa gttcaggggc catttcctga tcgaggggga
3601 tctcaacccg gacaatagcg atgttgacaa gctgttcatc cagctcgtgc agacgtacaa
3661 ccagctcttc gaggagaacc ccattaatgc gtcaggcgtc gacgcgaagg ctatcctgtc
3721 cgctaggctc tcgaagtctc ggcgcctcga gaacctgatc gcccagctgc cgggcgagaa
3781 gaagaacggc ctgttcggga atctcattgc gctcagcctg gggctcacgc ccaacttcaa
3841 gtcgaatttc gatctcgctg aggacgccaa gctgcagctc tccaaggaca catacgacga
3901 tgacctggat aacctcctgg cccagatcgg cgatcagtac gcggacctgt tcctcgctgc
3961 caagaatctg tcggacgcca tcctcctgtc tgatattctc agggtgaaca ccgagattac
4021 gaaggctccg ctctcagcct ccatgatcaa gcgctacgac gagcaccatc aggatctgac
4081 cctcctgaag gcgctggtca ggcagcagct ccccgagaag tacaaggaga tcttcttcga
4141 tcagtcgaag aacggctacg ctgggtacat tgacggcggg gcctctcagg aggagttcta
4201 caagttcatc aagccgattc tggagaagat ggacggcacg gaggagctgc tggtgaagct
4261 caatcgcgag gacctcctga ggaagcagcg gacattcgat aacggcagca tcccacacca
4321 gattcatctc ggggagctgc acgctatcct gaggaggcag gaggacttct accctttcct
4381 caaggataac cgcgagaaga tcgagaagat tctgactttc aggatcccgt actacgtcgg
4441 cccactcgct aggggcaact cccgcttcgc ttggatgacc cgcaagtcag aggagacgat
4501 cacgccgtgg aacttcgagg aggtggtcga caagggcgct agcgctcagt cgttcatcga
4561 gaggatgacg aatttcgaca agaacctgcc aaatgagaag gtgctcccta agcactcgct
4621 cctgtacgag tacttcacag tctacaacga gctgactaag gtgaagtatg tgaccgaggg
4681 catgaggaag ccggctttcc tgtctgggga gcagaagaag gccatcgtgg acctcctgtt
4741 caagaccaac cggaaggtca cggttaagca gctcaaggag gactacttca agaagattga
4801 gtgcttcgat tcggtcgaga tctctggcgt tgaggaccgc ttcaacgcct ccctggggac
4861 ctaccacgat ctcctgaaga tcattaagga taaggacttc ctggacaacg aggagaatga
4921 ggatatcctc gaggacattg tgctgacact cactctgttc gaggaccggg agatgatcga
4981 ggagcgcctg aagacttacg cccatctctt cgatgacaag gtcatgaagc agctcaagag
5041 gaggaggtac accggctggg ggaggctgag caggaagctc atcaacggca ttcgggacaa
5101 gcagtccggg aagacgatcc tcgacttcct gaagagcgat ggcttcgcga accgcaattt
5161 catgcagctg attcacgatg acagcctcac attcaaggag gatatccaga aggctcaggt
5221 gagcggccag ggggactcgc tgcacgagca tatcgcgaac ctcgctggct cgccagctat
5281 caagaagggg attctgcaga ccgtgaaggt tgtggacgag ctggtgaagg tcatgggcag
5341 gcacaagcct gagaacatcg tcattgagat ggcccgggag aatcagacca cgcagaaggg
5401 ccagaagaac tcacgcgaga ggatgaagag gatcgaggag ggcattaagg agctggggtc
5461 ccagatcctc aaggagcacc cggtggagaa cacgcagctg cagaatgaga agctctacct
5521 gtactacctc cagaatggcc gcgatatgta tgtggaccag gagctggata ttaacaggct
5581 cagcgattac gacgtcgatg ccatcgttcc acagtcattc ctgaaggatg actccattga
5641 caacaaggtc ctcaccaggt cggacaagaa ccggggcaag tctgataatg ttccttcaga
5701 ggaggtcgtt aagaagatga agaactactg gcgccagctc ctgaatgcca agctgatcac
5761 gcagcggaag ttcgataacc tcacaaaggc tgagaggggc gggctctctg agctggacaa
5821 ggcgggcttc atcaagaggc agctggtcga gacacggcag atcactaagc acgttgcgca
5881 gattctcgac tcacggatga acactaagta cgatgagaat gacaagctga tccgcgaggt
5941 gaaggtcatc accctgaagt caaagctcgt ctccgacttc aggaaggatt tccagttcta
6001 caaggttcgg gagatcaaca attaccacca tgcccatgac gcgtacctga acgcggtggt
6061 cggcacagct ctgatcaaga agtacccaaa gctcgagagc gagttcgtgt acggggacta
6121 caaggtttac gatgtgagga agatgatcgc caagtcggag caggagattg gcaaggctac
6181 cgccaagtac ttcttctact ctaacattat gaatttcttc aagacagaga tcactctggc
6241 caatggcgag atccggaagc gccccctcat cgagacgaac ggcgagacgg gggagatcgt
6301 gtgggacaag ggcagggatt tcgcgaccgt caggaaggtt ctctccatgc cacaagtgaa
6361 tatcgtcaag aagacagagg tccagactgg cgggttctct aaggagtcaa ttctgcctaa
6421 gcggaacagc gacaagctca tcgcccgcaa gaaggactgg gatccgaaga agtacggcgg
6481 gttcgacagc cccactgtgg cctactcggt cctggttgtg gcgaaggttg agaagggcaa
6541 gtccaagaag ctcaagagcg tgaaggagct gctggggatc acgattatgg agcgctccag
6601 cttcgagaag aacccgatcg atttcctgga ggcgaagggc tacaaggagg tgaagaagga
6661 cctgatcatt aagctcccca agtactcact cttcgagctg gagaacggca ggaagcggat
6721 gctggcttcc gctggcgagc tgcagaaggg gaacgagctg gctctgccgt ccaagtatgt
6781 gaacttcctc tacctggcct cccactacga gaagctcaag ggcagccccg aggacaacga
6841 gcagaagcag ctgttcgtcg agcagcacaa gcattacctc gacgagatca ttgagcagat
6901 ttccgagttc tccaagcgcg tgatcctggc cgacgcgaat ctggataagg tcctctccgc
6961 gtacaacaag caccgcgaca agccaatcag ggagcaggct gagaatatca ttcatctctt
7021 caccctgacg aacctcggcg cccctgctgc tttcaagtac ttcgacacaa ctatcgatcg
7081 caagaggtac acaagcacta aggaggtcct ggacgcgacc ctcatccacc agtcgattac
7141 cggcctctac gagacgcgca tcgacctgtc tcagctcggg ggcgacaagc ggccagcggc
7201 gacgaagaag gcggggcagg cgaagaagaa gaagtccggg tcctcggggt cgccaaagaa
7261 gaagcggaag gtcggctcaa gcgggtcacc aaagaagaag cggaaggtcg gctcgatgga
7321 cgctaagtcc ctcaccgctt ggagccgcac cctggtgacg ttcaaggacg tgttcgtcga
7381 tttcacaagg gaggagtgga agctcctgga cactgctcag cagatcgttt accggaacgt
7441 gatgctggag aactacaaga acctcgtgtc cctcggctac cagctgacga agccggatgt
7501 tattctccgc ctggagaagg gcgaggagcc cgggtcccct aagaagaagc ggaaggtcgg
7561 ctcctaccca tacgatgtcc ctgactacgc ctcatgagct caggcctcca tacaagtatt
7621 ggggatccac tacgaatttc cccgatcgtt caaacatttg gcaataaagt ttcttaagat
7681 tgaatcctgt tgccggtctt gcgatgatta tcatataatt tctgttgaat tacgttaagc
7741 atgtaataat taacatgtaa tgcatgacgt tatttatgag atgggttttt atgattagag
7801 tcccgcaatt atacatttaa tacgcgatag aaaacaaaat atagcgcgca aactaggata
7861 aattatcgcg cgcggtgtca tctatgttac tagatcggaa ttcgaggcgg tttgcgtatt
7921 ggctagagca gcttgccaac atggtggagc acgacactct cgtctactcc aagaatatca
7981 aagatacagt ctcagaagac caaagggcta ttgagacttt tcaacaaagg gtaatatcgg
8041 gaaacctcct cggattccat tgcccagcta tctgtcactt catcaaaagg acagtagaaa
8101 aggaaggtgg cacctacaaa tgccatcatt gcgataaagg aaaggctatc gttcaagatg
8161 cctctgccga cagtggtccc aaagatggac ccccacccac gaggagcatc gtggaaaaag
8221 aagacgttcc aaccacgtct tcaaagcaag tggattgatg tgataacatg gtggagcacg
8281 acactctcgt ctactccaag aatatcaaag atacagtctc agaagaccaa agggctattg
8341 agacttttca acaaagggta atatcgggaa acctcctcgg attccattgc ccagctatct
8401 gtcacttcat caaaaggaca gtagaaaagg aaggtggcac ctacaaatgc catcattgcg
8461 ataaaggaaa ggctatcgtt caagatgcct ctgccgacag tggtcccaaa gatggacccc
8521 cacccacgag gagcatcgtg gaaaaagaag acgttccaac cacgtcttca aagcaagtgg
8581 attgatgtga tatctccact gacgtaaggg atgacgcaca atcccactat ccttcgcaag
8641 accttcctct atataaggaa gttcatttca tttggagagg acacgctgaa atcaccagtc
8701 tctctctaca aatctatctc tctcgagctt tcgcagatcc cggggggcaa tgagatatga
8761 aaaagcctga actcaccgcg acgtctgtcg agaagtttct gatcgaaaag ttcgacagcg
8821 tctccgacct gatgcagctc tcggagggcg aagaatctcg tgctttcagc ttcgatgtag
8881 gagggcgtgg atatgtcctg cgggtaaata gctgcgccga tggtttctac aaagatcgtt
8941 atgtttatcg gcactttgca tcggccgcgc tcccgattcc ggaagtgctt gacattgggg
9001 agtttagcga gagcctgacc tattgcatct cccgccgtgc acagggtgtc acgttgcaag
9061 acctgcctga aaccgaactg cccgctgttc tacaaccggt cgcggaggct atggatgcga
9121 tcgctgcggc cgatcttagc cagacgagcg ggttcggccc attcggaccg caaggaatcg
9181 gtcaatacac tacatggcgt gatttcatat gcgcgattgc tgatccccat gtgtatcact
9241 ggcaaactgt gatggacgac accgtcagtg cgtccgtcgc gcaggctctc gatgagctga
9301 tgctttgggc cgaggactgc cccgaagtcc ggcacctcgt gcacgcggat ttcggctcca
9361 acaatgtcct gacggacaat ggccgcataa cagcggtcat tgactggagc gaggcgatgt
9421 tcggggattc ccaatacgag gtcgccaaca tcttcttctg gaggccgtgg ttggcttgta
9481 tggagcagca gacgcgctac ttcgagcgga ggcatccgga gcttgcagga tcgccacgac
9541 tccgggcgta tatgctccgc attggtcttg accaactcta tcagagcttg gttgacggca
9601 atttcgatga tgcagcttgg gcgcagggtc gatgcgacgc aatcgtccga tccggagccg
9661 ggactgtcgg gcgtacacaa atcgcccgca gaagcgcggc cgtctggacc gatggctgtg
9721 tagaagtact cgccgatagt ggaaaccgac gccccagcac tcgtccgagg gcaaagaaat
9781 agagtagatg ccgaccggat ctgtcgatcg acaagctcga gtttctccat aataatgtgt
9841 gagtagttcc cagataaggg aattagggtt cctatagggt ttcgctcatg tgttgagcat
9901 ataagaaacc cttagtatgt atttgtattt gtaaaatact tctatcaata aaatttctaa
9961 ttcctaaaac caaaatccag tactaaaatc cagatccccc gaattaattc ggcgttaatt
10021 cagtacatta aaaacgtccg caatgtgtta ttaagttgtc actagtcagg ttaactcaat
10081 tcggcgttaa ttcagtacat taaaaacgtc cgcaatgtgt tattaagttg tctaagcgtc
10141 aatttgttta caccacaata tatcctgcca ccagccagcc aacagctccc cgaccggcag
10201 ctcggcacaa aatcaccact cgatacaggc agcccatcag tccgggacgg cgtcagcggg
10261 agagccgttg taaggcggca gactttgctc atgttaccga tgctattcgg aagaacggca
10321 actaagctgc cgggtttgaa acacggatga tctcgcggag ggtagcatgt tgattgtaac
10381 gatgacagag cgttgctgcc tgtgatcact taagtaacta actaacagga agagtttgta
10441 gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt agtttgatgc ctggcagttt
10501 atggcgggcg tcctgcccgc caccctccgg gccgttgctt cacaacgttc aaatccgctc
10561 ccggcggatt tgtcctactc aggagagcgt tcaccgacaa acaacagata aaacgaaagg
10621 cccagtcttc cgactgagcc tttcgtttta tttgatgcct ggcagttccc tactctcgct
10681 tagtagttag acgtccccga gatccatgct agaccatgaa tccagaagcc cgagaggttg
10741 ccgcctttcg ggctttttct ttttcaaaaa aaaaaattta taaaacgatc tgttgcggcc
10801 ggccgccggg ttgtgggcaa aggcgctcga cggtgggcaa ccgcttgcgg ttgtccacgg
10861 gcggagccgg tgcgcgtagc gcattgtcca caagccaagg gcgaccaata attgatatat
10921 atattcataa ttgaaaagct aattgaacat actacttgct gtaactactt gccggagcga
10981 ggggtgtttg caagctgttg atctgaaagg gctattagcg ttctcacgtg cctttttgat
11041 tagcgatttc acgtgacctt attagcgatt tcacgtactc cgattagcga tttcacgtac
11101 cctgattagc gatttcacgt ggatagtttt tggagcgggc cggaaagccc cgtgaatcaa
11161 ggctttgcgg ggcattagcg gtttcacgtg gataactacc ctctatccac aggcttccgg
11221 ggataaaaaa gcccgctcga cggcgggctg ttggatgggg atctagcggt aatacggtta
11281 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
11341 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
11401 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
11461 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
11521 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
11581 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
11641 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
11701 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
11761 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
11821 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
11881 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
11941 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctaccgggtc tgacgctcag
12001 tggaacgggg cccaatctga ataatgttac aaccaattaa ccaattctga ttagaaaaac
12061 tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt
12121 tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca
12181 agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc
12241 ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt
12301 gagaatggca aaagtttatg catttctttc cagacttgtt caacaggcca gccattacgc
12361 tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg
12421 agacgaaata cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg
12481 cgcagggaca ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat
12541 acctggaatg ctgtttttcc ggggatcgca gtggtgagta accatgcatc atcaggagta
12601 cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc
12661 atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc
12721 gcatcgggct tcccatacaa gcgatagatt gtcgcacctg attgcccgac attatcgcga
12781 gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg cctcgacgtt
12841 tcccgttgaa tatggctcat aacacccctt gtattactgt ttatgtaagc agacagtttt
12901 attgttcatg atgatatatt tttatcttgt gcaatgtaac atcagagatt ttgagacacg
12961 ggccagagct gcagtttgat cccgagggga accctgtggt tgacatgcac atacaaatgg
13021 acgaacggat aaaccttttc acgccctttt aaatatccgt tattctaata aacgctcttt
13081 tctcttag
//
